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Luciferase mRNA Transfection of Antigen Presenting Cells Permits Sensitive Nonradioactive Measurement of Cellular and Humoral Cytotoxicity
Joint Authors
Türeci, Özlem
Omokoko, Tana A.
Luxemburger, Uli
Bardissi, Shaheer
Simon, Petra
Utsch, Magdalena
Breitkreuz, Andrea
Sahin, Ugur
Source
Journal of Immunology Research
Issue
Vol. 2016, Issue 2016 (31 Dec. 2016), pp.1-13, 13 p.
Publisher
Hindawi Publishing Corporation
Publication Date
2016-01-05
Country of Publication
Egypt
No. of Pages
13
Main Subjects
Abstract EN
Immunotherapy is rapidly evolving as an effective treatment option for many cancers.
With the emerging fields of cancer vaccines and adoptive cell transfer therapies, there is an increasing demand for high-throughput in vitro cytotoxicity assays that efficiently analyze immune effector functions.
The gold standard 51Cr-release assay is very accurate but has the major disadvantage of being radioactive.
We reveal the development of a versatile and nonradioactive firefly luciferase in vitro transcribed (IVT) RNA-based assay.
Demonstrating high efficiency, consistency, and excellent target cell viability, our optimized luciferase IVT RNA is used to transfect dividing and nondividing primary antigen presenting cells.
Together with the long-lasting expression and minimal background, the direct measurement of intracellular luciferase activity of living cells allows for the monitoring of killing kinetics and displays paramount sensitivity.
The ability to cotransfect the IVT RNA of the luciferase reporter and the antigen of interest into the antigen presenting cells and its simple read-out procedure render the assay high-throughput in nature.
Results generated were comparable to the 51Cr release and further confirmed the assay’s ability to measure antibody-dependent cell-mediated cytotoxicity and complement-dependent cytotoxicity.
The assay’s combined simplicity, practicality, and efficiency tailor it for the analysis of antigen-specific cellular and humoral effector functions during the development of novel immunotherapies.
American Psychological Association (APA)
Omokoko, Tana A.& Luxemburger, Uli& Bardissi, Shaheer& Simon, Petra& Utsch, Magdalena& Breitkreuz, Andrea…[et al.]. 2016. Luciferase mRNA Transfection of Antigen Presenting Cells Permits Sensitive Nonradioactive Measurement of Cellular and Humoral Cytotoxicity. Journal of Immunology Research،Vol. 2016, no. 2016, pp.1-13.
https://search.emarefa.net/detail/BIM-1108921
Modern Language Association (MLA)
Omokoko, Tana A.…[et al.]. Luciferase mRNA Transfection of Antigen Presenting Cells Permits Sensitive Nonradioactive Measurement of Cellular and Humoral Cytotoxicity. Journal of Immunology Research No. 2016 (2016), pp.1-13.
https://search.emarefa.net/detail/BIM-1108921
American Medical Association (AMA)
Omokoko, Tana A.& Luxemburger, Uli& Bardissi, Shaheer& Simon, Petra& Utsch, Magdalena& Breitkreuz, Andrea…[et al.]. Luciferase mRNA Transfection of Antigen Presenting Cells Permits Sensitive Nonradioactive Measurement of Cellular and Humoral Cytotoxicity. Journal of Immunology Research. 2016. Vol. 2016, no. 2016, pp.1-13.
https://search.emarefa.net/detail/BIM-1108921
Data Type
Journal Articles
Language
English
Notes
Includes bibliographical references
Record ID
BIM-1108921